iCell® CardiomyocytesBasic cardiac research, drug discovery, and toxicity

testing require an in vitro system that recapitulates

native biology. FUJIFILM Cellular Dynamics, Inc. (FCDI),

meets this need with iCell® Cardiomyocytes, human

cardiomyocytes with proven, predictive, and published

advantages over other cellular cardiac models for

faithfully and robustly reflecting cardiovascular disease

endpoints and for enabling early and accurate prediction

of drug-induced cardiotoxicity. The human biology,

ease of use, and far-reaching applicability of iCell

Cardiomyocytes has had a profound impact on academic

research and the search for new medicines and cures.

Genomic and Protein Characterization

iCell Cardiomyoctyes are differentiated from human

induced pluripotent stem cells. Whole-genome transcript

profiling and immunocytochemical characterization

demonstrate that iCell Cardiomyocytes exhibit a stable

human cardiac gene expression profile with proper

protein expression and localization necessary for cardiac

function.

Human cells: iCell Cardiomyocytes are terminally differentiated from human iPS cells and exhibit functional characteristics similar to native human cardiomyocytes.

Homogenous and reproducible: iCell Cardiomyocytes are highly pure, providing biologically relevant and reproducible results.

Acute and long-term testing: iCell Cardiomyocytes remain viable and pure in culture for weeks, enabling assessment of both acute and sub-chronic responses.

Easy to implement: iCell Cardiomyocytes are shipped cryopreserved with cell culture media specifically formulated for optimal cell performance. Simply thaw and use.

Advantages

p Figure 1: Stable and Appropriate Gene Expression Data for 203 cardiac genes demonstrate a stable genomic expression profile for iCell Cardiomyocytes that trends well with that of adult human cardiac tissue. (Data were provided by Hoffmann-La Roche and adapted from Babiarz et al. (2011) Stem Cells and Dev.)*

p Figure 2: Pure Pan-cardiomyocyte Population (A) Flow cytometry (cTNT, a-Actinin gating) demonstrates the >97% purity of iCell Cardiomyocytes. (B, C) Immunocytochemical labeling shows a pan-cardiac population with a sarcomeric myofibril organization. (Data were adapted from Kattman et al. (2011) J Cardiovasc Transl Res.)*

Ca-Actinin / Hoechst

BcTNT / MLC2V / Hoechst

A

cTNT

a-Actinin

0.47% 97.42%

0.10% 2.02%

Adult Human Cardiac Tissue

iCell Cardiomyocytes at End of DifferentiationiCell Cardiomyocytesat ~90 Days Post-differentiation

PRODUCT DATASHEET

Electrophysiological Characterization

experiments demonstrate that the electrophysiological

properties of iCell Cardiomyocytes are similar to native

human cardiac myocytes.

t Figure 3: Action Potential RecordingSpontaneous action potentials are classified as atrial-, nodal-, and ventricular-like, with ~50% of the iCell Cardiomyocytes showing a ventricular-like action potential phenotype. (Data were adapted from Ma et al. (2011) Am J Physiol Heart Circ Physiol.)*

Atrial-like Nodal-like Ventricular-like

0 mV

20 mV

200 ms

iCell Cardiomyocytes are electrically active with

spontaneously occurring action potentials driving rhythmic

contractions. High precision current- and voltage-clamp

Toxicity Testing

Cardiomyocytes especially well suited for discriminating

between generalized and cell-specific toxicities.

p Figure 5: Cell-specific Effects(A) General toxicants demonstrate adverse effects on iCell Cardiomyocytes. (B, C) The HDAC inhibitor Vorinostat (SAHA) shows cell-type specific toxicity. iCell Cardiomyocytes were unaffected by compound application whereas the commonly used immortalized K562 cell line showed clear toxicity. Using only immortalized cell lines in such cell-based assays could result in false positive or negative assessments of terminally differentiated cell toxicity. (Data were provided by Promega Corp.)*

20000

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-10 -9 -8 -7 -6 -5Log [SAHA] M

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Luminescence (R

LU)

Cytotoxicity EC50 = ND

Viability EC50 = ND

Caspase EC50 = ND

Cytotoxicity EC50 = 4.5 µM

Viability EC50 = 1.7 µM

Caspase EC50 = 3.3 µM

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Doxorubicin

Doxazosin mesylate

Valinomycin

Gleevec

Isoproterenol

iCell CardiomyocytesA B

K562C

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The adherent nature, ease of use, and recapitulation

of human in vivo cardiac myocyte behavior make iCell

t Figure 4: Ionic CurrentsiCell Cardiomyocytes contain the (A) expected human cardiac ionic currents and show the (B) expected effects when exposed to ion channel-blocking drugs. (Data were adapted from Ma et al. (2011) Am J Physiol Heart Circ Physiol.)*

ICa-L

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ICa Block (shortened APD)

Control

Nifedipine 3 nM

Nifedipine 10 nMNifedipine 30 nM

Nifedipine 100 nM

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ControlE4031 3 nM

E4031 10 nME4031 30 nM

E4031 100 nM

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EADGeneration

IKr Block (prolonged APD)B

100 pA500 ms

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IfunnyIK1Control

BaCl2 Subtraction

500 µM BaCl2

Voltage (mV)

-73 mV-123 mV 2 s

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nt De

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EETHigher Throughput Functional Testing

Reporter Assays and Microscopy Applications

t Figure 6: Higher Throughput Functional Analysis (A) Effects of ion channel and GPCR modulation on beating behavior were assayed using the xCELLigence RTCA Cardio System (Roche Applied Science). (B) Metabolic profiling with different energy substrates were assayed using the XF96 Extracellular Flux Analyzer (Seahorse Bioscience). (C) GPCR-mediated changes in Ca2+ cycling and contraction rate were assayed using the FLIPR® Tetra System (Molecular Devices, Inc.). (Data were provided by Molecular Devices, Inc.)*

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Log [Isoproterenol] µM

Beat

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0 6 13 19 25 32 38 44 51 57Time (min)

241214187160133106805326-1

-28

OCR

pM

ole/

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C400

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100 0 20 40 60 80 100

Time (sec)

RFU

Addition of Isoproterenol

AControl

E4031

examine compound effects in a relevant human cellular

environment early in the discovery process.

Their adherent nature and in vitro viability enable iCell

Cardiomyocytes to be used for live or fixed reporter assays and microscopy applications uniquely tailored

to an individual investigator’s requirements.

iCell Cardiomyocytes are suitable for examining

functional endpoints across many higher throughput

platforms, thus affording investigators the opportunity to

t Figure 7: Reporter Assays and Microscopy Applications (A) GFP transiently transfected into iCell Cardiomyocytes verifies relatively high transfection efficiencies. (B) Isoproterenol induces a high level of transiently transfected CRE-luciferase activity (note log scale). (C) Fluorescent microscopy image illustrates the perinuclear localization of BNP staining. (D) Endothelin-induced increases in BNP expression were measured by fluorescent microscopy.*

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Log ET-1 (M)

Field

Rela

tve In

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iCell Products

Provide access to biologically relevant, human iPS cells for disease modeling, drug discovery, toxicity testing, and regenerative medicine. FCDI’s rapidly growing portfolio of iCell products includes human cardiomyocytes, GABAergic, glutamatergic, dopaminergic and motor neurons, hepatocytes, endothelial cells, astrocytes, hematopoietic progenitor cells, skeletal myoblasts, macrophages, and others.

Visit the FCDI website for the most current list of supported cell types.

DS-CM180817 © Copyright 2018 FUJIFILM Cellular Dynamics, Inc.

Applications

iCell Cardiomyocytes are amenable to use on multiple platforms to

study a variety of cellular functions and assay endpoints including:

Proarrhythmia detection Biochemical and structural toxicity Bioenergetics Contractility Disease modeling Phenotypic screening Target identification Hit-to-lead validation and lead optimization

For More Information

FUJIFILM Cellular Dynamics, Inc. 525 Science Drive Madison, WI 53711 USA

(608) 310-5100 | Toll-free US (877) 310-6688 fcdi-sales@fujifilm.com www.fujifilmdi.com

Specifications

Cell Type Cardiomyocytes

Organism Human

Source Differentiated from an FCDI reprogrammed human iPS cell line

Purity >95% cardiomyocytes

Quantity ≥1.0 x 106 or ≥4.0 x 106 viable cells per vial

Shipped Frozen

Ordering Information

Kit Component(s)1 Catalog Number

iCell Cardiomyocytes Kit, 01434

≥1.0 x 106 viable cells 30 ml Plating Medium 100 ml Maintenance Medium

R1057

≥4.0 x 106 viable cells 30 ml Plating Medium 2 x 100 ml Maintenance Medium

R1007

iCell Cardiomyocytes Kit, 11713

≥1.0 x 106 viable cells 30 ml Plating Medium 100 ml Maintenance Medium

R1105

3 x ≥1.0 x 106 viable cells 30 ml Plating Medium 2 x 100 ml Maintenance Medium

R1117

≥4.0 x 106 viable cells 30 ml Plating Medium 2 x 100 ml Maintenance Medium

R1106

iCell Cardiomyocytes Maintenance Medium

100 ml Maintenance Medium M1003

1 A User's Guide is provided in each kit.

* Images and data are representative of iCell Cardiomyocytes derived from two different genetic backgrounds (donors 01434 and 11713).